Specific and behaviorally consequential astrocyte G_q GPCR signaling attenuation in vivo with iβARK
Abstract
Astrocytes respond to neurotransmitters and neuromodulators using G-protein-coupled receptors (GPCRs) to mediate physiological responses. Despite their importance, there has been no method to genetically, specifically, and effectively attenuate astrocyte G_q GPCR pathways to explore consequences of this prevalent signaling mechanism in vivo. We report a 122-residue inhibitory peptide from β-adrenergic receptor kinase 1 (iβARK; and inactive D110A control) to attenuate astrocyte G_q GPCR signaling. iβARK significantly attenuated G_q GPCR Ca²⁺ signaling in brain slices and, in vivo, altered behavioral responses, spared other GPCR responses, and did not alter astrocyte spontaneous Ca²⁺ signals, morphology, electrophysiological properties, or gene expression in the striatum. Furthermore, brain-wide attenuation of astrocyte G_q GPCR signaling with iβARK using PHP.eB adeno-associated viruses (AAVs), when combined with c-Fos mapping, suggested nuclei-specific contributions to behavioral adaptation and spatial memory. iβARK extends the toolkit needed to explore functions of astrocyte G_q GPCR signaling within neural circuits in vivo.
Additional Information
© 2021 Elsevier Inc. Received 20 November 2020, Revised 14 April 2021, Accepted 18 May 2021, Available online 16 June 2021. We thank the UCLA Neuroscience Genomics Core for assistance with sequencing and Fuying Gao for helping with data analysis. Thanks to UCLA Behavioral Testing Core for guidance and equipment. Collaborations between the B.S.K. and V.G. groups are supported by National Institutes of Health (NIH) grant DA047444. This work was supported by NIH grant R35NS111583, an Allen Distinguished Investigator Award through the Paul G. Allen Frontiers Group, and the Ressler Family Foundation (B.S.K.). S.M.S. was supported by the Brody Family Medical Trust Fund Fellowship and NIH grant HL132882. J.N. was partly supported by a JSPS Overseas Research Fellowship (H28-729) and the Uehara Memorial Foundation Overseas Postdoctoral Research Fellowship (201730082). X.Y. was partly supported by the American Heart Association (16POST27260256). We acknowledge support from the NINDS Informatics Center for Neurogenetics and Neurogenomics (grant P30 NS062691 to G.C.) and the Genetics, Genomics and Informatics Core of the Semel Institute of Neuroscience at UCLA (grant U54HD087101-01 from the Eunice Kennedy Shriver National Institute of Child Health and Human Development). P.G. and A.B. were supported by NIH grant R01NS116589. V.G. was supported by a NIH Pioneer Award (DP1OD025535), the Vallee Foundation, the CZI Neurodegeneration Challenge Network, and the Beckman Institute for CLARITY, Optogenetics and Vector Engineering Research (CLOVER) for technology development and dissemination. Author contributions: J.N. carried out most of the experiments, including molecular biology, mouse surgeries, IHC, slice imaging, RNA-seq, and behavior, with help from X.Y. and B.D.-C. X.Y. performed analyses of RNA-seq. A.B. performed in vivo cortical calcium imaging experiments, including those surgeries. Z.Q. generated the AAV-PHP.eB viruses with guidance from V.G., who provided equipment. S.M.S. discussed iβARK with B.S.K. at the germinal stages and shared reagents and insights. M.O. and M.R.G. helped with revision experiments. G.C. provided guidance on the analysis of RNA-seq data. P.G. supervised the in vivo imaging experiments. B.S.K. conceived, designed, and directed the project; guided analyses; and performed electrophysiology. B.S.K. and J.N. wrote the paper, and all authors commented. The authors declare no competing interests relevant to this study. However, B.S.K. is a consultant for Third Rock Ventures.Attached Files
Accepted Version - nihms-1716140.pdf
Supplemental Material - 1-s2.0-S0896627321003767-mmc1.pdf
Supplemental Material - 1-s2.0-S0896627321003767-mmc2.xlsx
Supplemental Material - 1-s2.0-S0896627321003767-mmc3.mp4
Supplemental Material - 1-s2.0-S0896627321003767-mmc4.mp4
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Additional details
- Alternative title
- Specific and behaviorally consequential astrocyte Gq GPCR signaling attenuation in vivo with iβARK
- PMCID
- PMC8418870
- Eprint ID
- 109586
- Resolver ID
- CaltechAUTHORS:20210625-220915439
- NIH
- DA047444
- NIH
- R35NS111583
- Paul G. Allen Family Foundation
- Ressler Family Foundation
- Brody Family Medical Trust Fund
- NIH
- HL132882
- Japan Society for the Promotion of Science (JSPS)
- H28-729
- Uehara Memorial Foundation
- 201730082
- American Heart Association
- 16POST27260256
- NIH
- P30 NS062691
- UCLA
- NIH
- U54HD087101-01
- NIH
- R01NS116589
- NIH
- DP1OD025535
- Vallee Foundation
- CZI Neurodegeneration Challenge Network
- Caltech Beckman Institute
- Created
-
2021-06-25Created from EPrint's datestamp field
- Updated
-
2023-07-07Created from EPrint's last_modified field
- Caltech groups
- Division of Biology and Biological Engineering (BBE)